GST-Tagged protein

hiiiiii,
I hav a problem in cutting GST-Tag from my protein.After incubating GST-tagged beads with precision protease for 16 hours, theoretically I should get a clean single band. But in my coomassie gel there are multiple bands.In western I am geting one more extra band below the size of my protein.How will I get a single band????

hiiiiii, I hav a problem in cutting GST-Tag from my protein.After incubating GST-tagged beads with precision protease for 16 hours, theoretically I should get a clean single band. But in my coomassie gel there are multiple bands.In western I am geting one more extra band below the size of my protein.How will I get a single band????

Yeah...even I think its degradation of protein....what temperature did you inubate it at for 16 hours?

This may be killing it.

Best,TC

hiiiiii, I hav a problem in cutting GST-Tag from my protein.After incubating GST-tagged beads with precision protease for 16 hours, theoretically I should get a clean single band. But in my coomassie gel there are multiple bands.In western I am geting one more extra band below the size of my protein.How will I get a single band????

Yeah...even I think its degradation of protein....what temperature did you inubate it at for 16 hours?

This may be killing it.

Best,TC

hiiiiii, I hav a problem in cutting GST-Tag from my protein.After incubating GST-tagged beads with precision protease for 16 hours, theoretically I should get a clean single band. But in my coomassie gel there are multiple bands.In western I am geting one more extra band below the size of my protein.How will I get a single band????